Abstract
Lentiviral systems have proven advantageous in the delivery and long-term integration of gene sequences into the genome of several cell types in vitro, in vivo, as well as in clinical trials. Here we detail the protocols involved in the molecular cloning of ADAMTSL2 and ADAMTSL4 into the human immunodeficiency virus (HIV)-derived pCDH lentiviral system. We also describe the lentiviral transduction of ADAMTSL2 and ADAMTSL4 into mammalian HEK293-EBNA cells to create stable cell lines, as well as their recombinant expression.
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Acknowledgments
MS was supported by a Medical Research Council (MRC) grant (ref: MR/L016540/1) and a Biotechnology and Biological Sciences Research Council (BBSRC) studentship to CB. The Wellcome Centre for Cell-Matrix Research is supported by core grant funding from the Wellcome Trust (088785/Z/09/Z). SAC is supported by a BBSRC grant (ref: BB/R008221/1) to CB.
We would like to extend our gratitude to the Flow Cytometry Facility (Faculty of Biology, Medicine and Health, University of Manchester, UK) for their assistance and expertise.
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Singh, M., Cain, S.A., Baldock, C. (2020). Molecular Cloning, Lentiviral Transduction, and Expression of Recombinant ADAMTSL2 and ADAMTSL4. In: Apte, S. (eds) ADAMTS Proteases. Methods in Molecular Biology, vol 2043. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9698-8_12
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DOI: https://doi.org/10.1007/978-1-4939-9698-8_12
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-4939-9698-8
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